Detection of Leptospira pomona in bovine semen by fluorescent capillary eletrophoresis

Detalhes bibliográficos
Autor(a) principal: Dias, Francisca Elda Ferreira
Data de Publicação: 2006
Outros Autores: Aoki, Sérgio Morais, Mesquita, Lígia Garcia, Nunes, Caris Maroni, Garcia, José Fernando
Tipo de documento: Artigo
Idioma: por
Título da fonte: Brazilian Journal of Veterinary Research and Animal Science
Texto Completo: https://www.revistas.usp.br/bjvras/article/view/26488
Resumo: This study was performed in order to evaluate the detection limit of PCR with fluorescent capillary electrophoresis for Leptospira pomona diagnosis in bovine semen. Negative bovine semen samples were artificially contaminated with Leptospira pomona (10(0) to 10(7) bacteria/ml) and DNA was extracted by phenol/chloroform protocol. DNA fragments visualization was done by three electrophoresis methods: under UV light in 2% agarose gel, silver staining 8% polyacrylamide gel and fluorescent capillary electrophoresis. The detection limit of capillary electrophoresis for Leptospira pomona was 10²bacteria/ml. Under UV light, in 2% agarose gel, the detection limit was of 10(4) bacteria/ml while for silver stained 8% polyacrylamide gel it was 10² bacteria/ml. PCR with fluorescent capillary electrophoresis is an efficient and rapid diagnostic test for DNA detection of Leptospira in bovine semen and this can be an important tool for herd and semen sanitary control in artificial insemination centers.
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spelling Detection of Leptospira pomona in bovine semen by fluorescent capillary eletrophoresisDetecção de Leptospira pomona em sêmen bovino por eletroforese capilar fluorescenteSêmen BovinoLeptospiraPCREletroforese CapilarSemen BovineLeptospiraPCRCapillary EletrophoresisThis study was performed in order to evaluate the detection limit of PCR with fluorescent capillary electrophoresis for Leptospira pomona diagnosis in bovine semen. Negative bovine semen samples were artificially contaminated with Leptospira pomona (10(0) to 10(7) bacteria/ml) and DNA was extracted by phenol/chloroform protocol. DNA fragments visualization was done by three electrophoresis methods: under UV light in 2% agarose gel, silver staining 8% polyacrylamide gel and fluorescent capillary electrophoresis. The detection limit of capillary electrophoresis for Leptospira pomona was 10²bacteria/ml. Under UV light, in 2% agarose gel, the detection limit was of 10(4) bacteria/ml while for silver stained 8% polyacrylamide gel it was 10² bacteria/ml. PCR with fluorescent capillary electrophoresis is an efficient and rapid diagnostic test for DNA detection of Leptospira in bovine semen and this can be an important tool for herd and semen sanitary control in artificial insemination centers.Este estudo pretendeu avaliar o limiar de detecção da técnica de PCR aliada à eletroforese capilar para diagnóstico da Leptospira pomona em sêmen bovino. Doses inseminantes livres de patógenos foram contaminadas experimentalmente com Leptospira pomona em escalas que variavam de 10(0) a 10(7) bactérias/ml e submetidas à extração de DNA pelo método de fenol/clorofórmio. Após a reação de PCR, a visualização dos fragmentos foi realizada em três tipos de eletroforese: agarose 2% sob luz UV, acrilamida 8% corado com prata e eletroforese capilar fluorescente. A detecção de DNA de Leptospira pomona em sêmen bovino através de eletroforese capilar fluorescente foi possível a partir de concentração de 10² bactérias/ml. Nos métodos de eletroforese em agarose 2%, observou-se limite de detecção de 10(4) bactérias/ml e em gel de poliacrilamida 8% o limite de detecção foi de 10² bactérias/ml. A eletroforese capilar demonstrou ser uma alternativa eficaz e rápida na detecção de DNA de Leptospira em sêmen bovino podendo ser uma valiosa ferramenta para controle de qualidade do sêmen produzido em centrais de inseminação artificial dada a facilidade de automação desse processo.Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia2006-06-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://www.revistas.usp.br/bjvras/article/view/2648810.11606/issn.1678-4456.bjvras.2006.26488Brazilian Journal of Veterinary Research and Animal Science; Vol. 43 Núm. 3 (2006); 394-399Brazilian Journal of Veterinary Research and Animal Science; Vol. 43 No. 3 (2006); 394-399Brazilian Journal of Veterinary Research and Animal Science; v. 43 n. 3 (2006); 394-399Brazilian Journal of Veterinary Research and Animal Science; V. 43 N. 3 (2006); 394-3991678-44561413-9596reponame:Brazilian Journal of Veterinary Research and Animal Scienceinstname:Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP)instacron:USPporhttps://www.revistas.usp.br/bjvras/article/view/26488/28271Dias, Francisca Elda FerreiraAoki, Sérgio MoraisMesquita, Lígia GarciaNunes, Caris MaroniGarcia, José Fernandoinfo:eu-repo/semantics/openAccess2020-06-23T04:17:48Zoai:revistas.usp.br:article/26488Revistahttps://www.revistas.usp.br/bjvrasPUBhttps://www.revistas.usp.br/bjvras/oaibjvras@usp.br1413-95961413-9596opendoar:https://www.revistas.usp.br/bjvras/index2023-01-12T16:42:46.095153Brazilian Journal of Veterinary Research and Animal Science - Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP)false
dc.title.none.fl_str_mv Detection of Leptospira pomona in bovine semen by fluorescent capillary eletrophoresis
Detecção de Leptospira pomona em sêmen bovino por eletroforese capilar fluorescente
title Detection of Leptospira pomona in bovine semen by fluorescent capillary eletrophoresis
spellingShingle Detection of Leptospira pomona in bovine semen by fluorescent capillary eletrophoresis
Dias, Francisca Elda Ferreira
Sêmen Bovino
Leptospira
PCR
Eletroforese Capilar
Semen Bovine
Leptospira
PCR
Capillary Eletrophoresis
title_short Detection of Leptospira pomona in bovine semen by fluorescent capillary eletrophoresis
title_full Detection of Leptospira pomona in bovine semen by fluorescent capillary eletrophoresis
title_fullStr Detection of Leptospira pomona in bovine semen by fluorescent capillary eletrophoresis
title_full_unstemmed Detection of Leptospira pomona in bovine semen by fluorescent capillary eletrophoresis
title_sort Detection of Leptospira pomona in bovine semen by fluorescent capillary eletrophoresis
author Dias, Francisca Elda Ferreira
author_facet Dias, Francisca Elda Ferreira
Aoki, Sérgio Morais
Mesquita, Lígia Garcia
Nunes, Caris Maroni
Garcia, José Fernando
author_role author
author2 Aoki, Sérgio Morais
Mesquita, Lígia Garcia
Nunes, Caris Maroni
Garcia, José Fernando
author2_role author
author
author
author
dc.contributor.author.fl_str_mv Dias, Francisca Elda Ferreira
Aoki, Sérgio Morais
Mesquita, Lígia Garcia
Nunes, Caris Maroni
Garcia, José Fernando
dc.subject.por.fl_str_mv Sêmen Bovino
Leptospira
PCR
Eletroforese Capilar
Semen Bovine
Leptospira
PCR
Capillary Eletrophoresis
topic Sêmen Bovino
Leptospira
PCR
Eletroforese Capilar
Semen Bovine
Leptospira
PCR
Capillary Eletrophoresis
description This study was performed in order to evaluate the detection limit of PCR with fluorescent capillary electrophoresis for Leptospira pomona diagnosis in bovine semen. Negative bovine semen samples were artificially contaminated with Leptospira pomona (10(0) to 10(7) bacteria/ml) and DNA was extracted by phenol/chloroform protocol. DNA fragments visualization was done by three electrophoresis methods: under UV light in 2% agarose gel, silver staining 8% polyacrylamide gel and fluorescent capillary electrophoresis. The detection limit of capillary electrophoresis for Leptospira pomona was 10²bacteria/ml. Under UV light, in 2% agarose gel, the detection limit was of 10(4) bacteria/ml while for silver stained 8% polyacrylamide gel it was 10² bacteria/ml. PCR with fluorescent capillary electrophoresis is an efficient and rapid diagnostic test for DNA detection of Leptospira in bovine semen and this can be an important tool for herd and semen sanitary control in artificial insemination centers.
publishDate 2006
dc.date.none.fl_str_mv 2006-06-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://www.revistas.usp.br/bjvras/article/view/26488
10.11606/issn.1678-4456.bjvras.2006.26488
url https://www.revistas.usp.br/bjvras/article/view/26488
identifier_str_mv 10.11606/issn.1678-4456.bjvras.2006.26488
dc.language.iso.fl_str_mv por
language por
dc.relation.none.fl_str_mv https://www.revistas.usp.br/bjvras/article/view/26488/28271
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia
publisher.none.fl_str_mv Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia
dc.source.none.fl_str_mv Brazilian Journal of Veterinary Research and Animal Science; Vol. 43 Núm. 3 (2006); 394-399
Brazilian Journal of Veterinary Research and Animal Science; Vol. 43 No. 3 (2006); 394-399
Brazilian Journal of Veterinary Research and Animal Science; v. 43 n. 3 (2006); 394-399
Brazilian Journal of Veterinary Research and Animal Science; V. 43 N. 3 (2006); 394-399
1678-4456
1413-9596
reponame:Brazilian Journal of Veterinary Research and Animal Science
instname:Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP)
instacron:USP
instname_str Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP)
instacron_str USP
institution USP
reponame_str Brazilian Journal of Veterinary Research and Animal Science
collection Brazilian Journal of Veterinary Research and Animal Science
repository.name.fl_str_mv Brazilian Journal of Veterinary Research and Animal Science - Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP)
repository.mail.fl_str_mv bjvras@usp.br
_version_ 1797051557718523904

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